A novel transfection method for mammalian cells using calcium alginate microbeads

The direct transfer of genetic materials into mammalian cells is an indispensable technique. We have developed calcium alginate (CA) microbeads which can deliver plasmid DNAs and yeast artificial chromosomes into plant and yeast cells. In this paper, we demonstrate the effective transfection of mammalian cells by CA microbeads immobilizing plasmid DNAs. The transfection was performed using the pEGFP-C1 plasmid containing the cytomegalovirus (CMV) promoter and enhanced green fluorescent protein (EGFP) gene. The transient expression of EGFP was observed 24 h after transfection. The expression efficiency was maximum when the concentration of sodium alginate was 1% and the amount of plasmid DNA was increased to 100 microg. The expression efficiency of our method using CA microbeads is 2-10 times higher than that of the polyethylene glycol (PEG) method. Our results suggest that the CA microbead mediated transfection of mammalian cells effectively delivers genetic materials into mammalian suspension cells.     

Development of new dosimetry using extended DNA fibers

We applied fluorescent microscopy to monitor the damage of DNA upon exposure to gamma radiation. Our developed dosimetry demonstrated that the number of breaks in DNA is proportional to the dose of the irradiation but is not dependent on dose rate of the irradiation and the GC content of DNA.     

Polynucleotides. XLV Synthesis and properties of poly(2'-azido-2'-deoxyinosinic acid)

Poly (2'-azido-2'-deoxyinosinic acid), [poly (Iz)], was synthesized from 2'-azido-2'-deoxyinosine diphosphate by the action of polynucleotide phosphorylase. Poly (Iz) has UV absorption properties similar to poly (I) and hypochromicity of 11% at 0.15M Na+ and neutrality. In solutions of high Na+ ion concentration, poly (Iz) forms a multi-stranded complex and its Tm at 1.0M Na+ ion concentration was 43 degrees. Upon mixing with poly (C), poly (Iz) forms a 1:1 complex having a Tm lower than that of poly (I)-poly (C) complex in the same conditions. The effect of substitution at the 2'-position of the poly (I) strand was discussed in relation to the interferon-inducing activity.     

Dynamic flying characteristics of an air bearing slider over a disk with grooves and distribution of material properties

Bit-patterned media (BPM) consisting of land parts for read/writing and grooves, which are almost filled with non-magnetic materials to prevent magnetic interference between recording bits, are considered to be promising media for achieving ultrahigh density recording. A flying head slider flying over a BPM disk suffers from variations in both the spacing and van der Waals (vdW) attractive forces, which induce slider vibrations and spacing fluctuations. In the present study, we considered that BPM disks not only have a groove depth distribution, but that they also have a distribution of material properties (e.g., refractive index), which gives rise to a distribution in the vdW force. In the dynamic responses of sliders with small groove depths and a small variation in the refractive index, spacing fluctuations are found to be a superposition of fluctuations due to slider behaviors (1) over a disk with transverse grooves in a uniform material with a uniform refractive index (Case 1) and (2) over a flat disk with a refractive index distribution (Case 2). When the effects of Cases 1 and 2 cancel each other, the spacing fluctuations for the two cases cancel each other, reducing the total spacing fluctuation.     

Deformation characteristics of ultra-thin liquid film considering temperature and film thickness dependence of surface tension

Thermocapillary deformations of an ultra-thin liquid film caused by temperature distribution were three-dimensionally analyzed using the unsteady and linearized long wave equation considering the temperature and film thickness dependence of surface tension. The temperature and film thickness dependence equation for the surface tension of a liquid was firstly established. The temperature dependence of the surface tension was obtained experimentally using a surface tensiometer and the film thickness dependence was obtained theoretically from the corrected van der Waals pressure equation for a symmetric multilayer system. Time evolutions of depression and groove of the ultra-thin liquid film caused by local heating were obtained quantitatively.     

Successive changes in community structure of an ethylbenzene-degrading sulfate-reducing consortium

The microbial community structure and successive changes in a mesophilic ethylbenzene-degrading sulfate-reducing consortium were for the first time clarified by the denaturing gradient gel electrophoresis (DGGE) analysis of the PCR amplified 16S rRNA gene fragments. At least ten bands on the DGGE gel were detected in the stationary phase. Phylogenetic analysis of the DGGE bands revealed that the consortium consisted of different eubacterial phyla including the delta subgroup of Proteobacteria, the order Sphingobacteriales, the order Spirochaetales, and the unknown bacterium. The most abundant band C was closely related to strain mXyS1, an m-xylene-degrading sulfate-reducing bacterium (SRB), and occurred as a sole band on DGGE gels in the logarithmic growth phase that 40% ethylbenzene was consumed accompanied by sulfide production. During further prolonged incubation, the dominancy of band C did not change. These results suggest that SRB corresponds to the most abundant band C and contributes mainly to the degradation of ethylbenzene coupled with sulfate reduction.     

Characterization of alternative plasticizers in poly(vinyl chloride) sheets for blood containers

This study aimed to optimize the ratio of dioctyl 4‐cyclohexene‐1,2‐dicarboxylate (DOTH) and di‐isononyl‐cyclohexane‐1,2‐dicarboxylate (DINCH^®) for use as plasticizers in poly(vinyl chloride) (PVC) sheets. We also evaluated the biological safety of DOTH for its potential to be part of a safe PVC‐based blood container. The suppression of hemolysis in mannitol‐adenine‐phosphate / red cell concentrates (MAP/RCC) with DOTH/(DINCH^®‐PVC) sheets and the elution of plasticizers from the sheets increased with higher DOTH compositions. The properties of the PVC sheet containing DOTH and DINCH^® in the ratio of 25:33 parts against PVC 100 parts as a weight were almost identical to the PVC sheet made of di(2‐ethylhexyl) phthalate. From a subchronic toxicity test, DOTH did not show any adverse effects on all organs, including the testes, epididymis, liver, and kidneys. The no‐observed‐adverse‐effect level was 300 mg/kg body weight/day in a rat. These results suggest that DOTH/DINCH^® (25:33) is a promising candidate for the replacement of di(2‐ethylhexyl) phthalate in blood containers. J. VINYL ADDIT. TECHNOL., 22:520–528, 2016. © 2015 Society of Plastics Engineers     

High performance 35 nm gate length CMOS with NO oxynitride gate dielectric and Ni salicide

The 35 nm gate length CMOS devices with oxynitride gate dielectric and Ni salicide have been fabricated to study the feasibility of higher performance operation. Nitrogen concentration in gate oxynitride was optimized to reduce gate current I/sub g/ and to prevent boron penetration in the pFET. The thermal budget in the middle of the line (MOL) process was reduced enough to realize shallower junction depth in the S/D extension regions and to suppress gate poly-Si depletion. Finally, the current drives of 676 /spl mu/A//spl mu/m in nFET and 272 /spl mu/A//spl mu/m in pFET at V/sub dd/=0.85 V (at I/sub off/=100 nA//spl mu/m) were achieved and they are the best values for 35 nm gate length CMOS reported to date.